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目的 :观察睫状神经营养因子 (CNTF)对皮质酮致原代培养海马神经元损伤的保护作用并探讨其可能机制。方法 :大鼠海马神经元原代培养 ,结晶紫法测定细胞存活率 ,TU NEL法检测细胞凋亡及应用免疫组化法 ,观察 CNTF对皮质酮致神经元存活率降低的保护作用 ,及其与 Bax和 Bcl- 2含量的关系。 结果 :皮质酮可剂量依赖地降低原代培养海马神经元的细胞存活率 (P<0 .0 1)。 1× 10 - 7m ol/ L的皮质酮作用 2 4h后 ,(1)约 45 %的海马神经元死亡 ;(2 )出现紫蓝色 TU NEL阳性细胞和圆形坏死细胞 ;(3) Bcl- 2免疫组化阳性神经元减少 ,着色浅淡 ;Bax免疫组化阳性神经元增加 ,着色深。同时给予培养细胞1× 10 - 7mol/ L 皮质酮和不同浓度的 CNTF2 4h后 ,发现与对照组相比 ,CNTF组原代培养海马神经元 (1)存活率升高 ;(2 )紫蓝色 TU NEL 阳性细胞和未着色的圆形坏死细胞均减少 ;(3) Bcl- 2免疫组化阳性神经元增加 ,着色深 ;Bax免疫组化阳性神经元减少 ,着色浅淡。结论 :凋亡和坏死过程共同参与了皮质酮致原代培养海马神经元损伤。外源性 CNTF可通过减少细胞坏死和凋亡而改善神经元损伤 ,其减轻细胞凋亡的作用可能是通过上调抑制凋亡蛋白 Bcl- 2和下调促进凋亡蛋白 Bax实现的
Objective: To observe the protective effect of ciliary neurotrophic factor (CNTF) on neuronal damage induced by corticosterone in primary cultured hippocampal neurons and its possible mechanism. Methods: Primary cultured hippocampal neurons were cultured, the cell viability was measured by crystal violet method, apoptosis was detected by TU NEL method and immunohistochemical method was used to observe the protective effect of CNTF on the decrease of corticosterone-induced neuronal survival rate And Bax and Bcl-2 content. Results: Corticosterone reduced the cell survival rate of primary cultured hippocampal neurons dose-dependently (P <0.01). (1) Approximately 45% of hippocampal neurons died after 1 × 10 ~ 7mol / L corticosterone treatment for 24 hours; (2) TUV NEL positive cells and circular necrotic cells appeared; (3) Bcl- 2 immunohistochemical positive neurons decreased, pale; Bax immunohistochemical positive neurons increased, deep color. At the same time, cultured cortisone at 1 × 10-7 mol / L and CNTF2 at different concentrations were cultured for 4 hours. Compared with the control group, the survival rate of primary cultured hippocampal neurons in CNTF group (1) was increased; (2) TUNEL positive cells and uncolored circular necrotic cells decreased; (3) Bcl-2 immunohistochemical positive neurons increased, deep color; Bax immunohistochemical positive neurons decreased, pale. Conclusion: The apoptotic and necrotic processes are involved in the cortical ketone-induced primary culture of hippocampal neurons. Exogenous CNTF can improve neuronal injury by reducing cell necrosis and apoptosis, and its effect of reducing apoptosis may be through up-regulating the apoptosis-inhibiting protein Bcl-2 and down-regulating the promoting apoptosis protein Bax